Abstract: Provided herein are methods for mapping immune response to an immunogen, comprising: immunizing a subject with an immunogen and obtaining sera from the immunized subject at multiple time intervals following immunization, wherein the sera comprises one or more immune complexes between the immunogen and serum antibodies; imaging, by electron microscopy, the sera obtained from the immunized subject in each of the time intervals, to obtain structural images of the one or more immune complexes formed between the immunogen and serum antibodies; mapping immune response to the immunogen by measuring differences in structural images obtained at different time intervals to simultaneously visualize diverse antibodies targeting distinct epitopes in the immunized subjects.

Abstract: Provided herein are methods for mapping antibody binding to an immunogen, comprising: immunizing a subject with an immunogen and obtaining sera from the immunized subject at multiple time intervals following immunization, wherein the sera comprises antibodies that are used to form one or more immune complexes with the immunogen; isolating the one or more immune complexes formed by the serum derived antibodies bound to the immunogen; imaging, by electron microscopy, the one or more immune complexes in each of the time intervals, to obtain structural images formed between the immunogen and serum antibodies; determining, from the plurality of structural images, immunogen-antibody binding site for each of the immune complexes obtained at the plurality of time intervals; mapping immunogen-antibody binding by measuring differences in structural images obtained at different time intervals to determine immunogen-antibody binding over multiple time intervals.

Abstract: Provided herein are methods for mapping immune response to an immunogen, comprising: immunizing a subject with an immunogen and obtaining sera from the immunized subject at multiple time intervals following immunization, wherein the sera comprises one or more immune complexes between the immunogen and serum antibodies; imaging, by electron microscopy, the sera obtained from the immunized subject in each of the time intervals, to obtain structural images of the one or more immune complexes formed between the immunogen and serum antibodies; mapping immune response to the immunogen by measuring differences in structural images obtained at different time intervals to simultaneously visualize diverse antibodies targeting distinct epitopes in the immunized subjects.

Abstract: The invention relates to a method for isolating a polypeptide reactive to influenza A hemagglutinin, comprising the steps of preparing a library of nucleic acid sequences, wherein each member of said library is attached to a polypeptide sequence encoded thereby, selecting members of said library by contacting the members with a panning antigen, wherein the panning antigen comprises a hemagglutinin stem region polypeptide sequence, subsequently removing members unreactive to the panning antigen, and selecting a member of said library by determining its binding to at least 5 influenza A hemagglutinin subtypes of subtype group 1 and 2. The invention further relates to polypeptides, particularly antibodies, obtainable by this method, that neutralize at least five influenza A subtypes of subtype group 1 and 2, to vaccines and uses thereof.

Abstract: The invention relates to a method for isolating a polypeptide reactive to influenza A hemagglutinin, comprising the steps of preparing a library of nucleic acid sequences, wherein each member of said library is attached to a polypeptide sequence encoded thereby, selecting members of said library by contacting the members with a panning antigen, wherein the panning antigen comprises a hemagglutinin stem region polypeptide sequence, subsequently removing members unreactive to the panning antigen, and selecting a member of said library by determining its binding to at least 5 influenza A hemagglutinin subtypes of subtype group 1 and 2. The invention further relates to polypeptides, particularly antibodies, obtainable by this method, that neutralize at least five influenza A subtypes of subtype group 1 and 2, to vaccines and uses thereof.